DE112013001457T5 - Transdermal application system - Google Patents

Abstract

A transdermal application system for transdermal drug administration is presented in the form of a transdermal release patch for the administration of insulin. The plaster contains cross-linked, amidated low methoxylated pectin, insulin and an active ingredient for better transfer.

Description

THIS INVENTION relates to a transdermal delivery system. It relates more particularly to transdermal delivery systems for the transdermal delivery of insulin.

Currently, about 135 million people suffer from diabetes mellitus. Given the projected increases (170%) in developing countries and (40%) in developed countries, this figure is expected to rise to 300 million by 2025 [1]. Insulin is the standard pharmaceutical compound used to treat diabetes and insulin is normally given as follows; intravenous (iv), intramuscular (im) or subcutaneous (sc), the route of injection being the most common route of administration. However, needle phobia and stress symptoms caused by multiple daily injections lead to discomfort and inconvenience, thereby creating the need for a less stressful method of administering insulin [2].

Transdermal delivery systems provide slow and controlled delivery of drugs, avoid metabolic metabolic disorders, maintain constant blood levels for prolonged periods, and reduce side effects, thereby increasing acceptance. Since 1990, many studies have been carried out to improve the transdermal administration of insulin [3-8]. Methods that have proven useful include electroporation [3], lipid-enriched electroporation [3], topically applied biphasic vesicles [4], easily deformable carriers [5], ultrasound [6] and microneedles [7]. An investigation has also shown the need for effective skin preparation and electrical reinforcement [8]. However, all of these studies have made use of either chemical permeators or electrical pulses and sound waves to enhance the administration and none of the methods used has used exclusively a transdermal patch for administration. The Applicant has now found that pectin can be used for the transdermal administration of insulin.

According to a first aspect of the invention there is provided a transdermal delivery system for the transdermal delivery of insulin, said device comprising in the form of a patch cross-linked amidated low methylated pectin, insulin and a transdermal delivery enhancer.

The enhancer of transdermal delivery may be dimethyl sulfoxide, sodium oleate, or sodium dodecyl sulfate (SDS or NaDS). The patch may contain an antioxidant. Further, it may contain an antibiotic.

The antioxidant may be vitamin E, optionally combined with eucalyptus oil.

The antibiotic may be puromycin.

The low-methoxylated pectin may have a degree of methoxylation between about 19 and 23 and a degree of amidation of about 24 to 31. Preferably, the degree of methoxylation is between 19 and 23 and the degree of amidation is between 24 and 31.

The transdermal patch may be in the form described.

According to a second aspect of the invention, there is provided a method of treating diabetes, the method comprising the direct application to the skin of a subject or animal of a transdermal patch comprising cross-linked, amidated, low-methoxylated pectin, insulin and a transdermal delivery enhancer provides for the delivery of insulin through the skin.

The patch can occur in the form described.

Thus, the invention provides a patch and method for delivery of insulin through the skin through the use of a small traditional medicated plaster. The active ingredient of the patch of the invention enables the direct transfer of insulin through the skin into the bloodstream. The transdermal delivery of insulin is known that high molecular weight drugs such as insulin are not readily able to penetrate the skin and therefore can not enter the bloodstream. The active ingredient of the patch of the invention overcomes this problem by the use of a chemical enhancer which facilitates the passage of native insulin through the skin. There are currently no approved insulin patches with transdermal delivery mechanisms for the administration of insulin.

The invention will now be described by way of example with reference to the following examples and figures

a schematic representation of two parts of a patch according to the invention;

the administration of the amidated pectin, insulin matrix patch of the invention; and

a comparison of oral glucose tolerance (OGT) responses of streptozotocin (STZ) -induced diabetes rats versus different doses of insulin in a pectin hydrogel patch using controlled laboratory animals; the values are presented as averages and vertical bars indicate SEM of the means (n = 6 in each group); * p <0.05 by comparison with the control animals; ** p <0.05 by comparison with all groups; A (control), B (low), C (int low), D (int high) and E (high).

shows an embodiment of the transdermal delivery system for administering the invention in the form of a transdermal patch 10 , The patch 10 has a rectangular shape and is 120 mm long and 100 mm wide. It contains a hydrofilm cover 12 with a centrally arranged gauze strip 14 which is 80 mm long and 50 mm wide, measured on the cover 12 , A circular gel body 16 with a diameter of 25 mm comprising cross-linked amidated low-methoxylated pectin having a degree of methoxylation of 23 and a degree of amidation of 24, human insulin, dimethyl sulfoxide and vitamin E is central to the gauze strip 14 arranged. The patch 10 comes with an adhesive cover 18 (not shown separately in the drawing) provided.

shows schematically the application of the plaster 10 on a rat 20 , The neck 22 the rat 20 is shaved smooth and the plaster 10 is applied to the shaved area. shows the plaster 10 secured and secured by a sheath 24 ,

EXAMPLES

Preparing insulin patches

Materials and methods

• Drugs: Biphasic insulin (Actraphane HM, Novo Nordisk, Canada) or human insulin (Isophan Human Insulin, Lilly France SA, Fegershiem).
• Matrix: Amidated, low-methoxylated pectin with a degree of methoxylation of 23 and a degree of amidation of 24.
• Cations for crosslinking: calcium chloride.
• Penetration enhancer: dimethyl sulfoxide.
• Adhesives: Self-adhesive bandages or hydrofilm (5 cm × 7.5 cm, 8 cm × 12 cm, 10 cm × 20 cm).
• Antioxidants: Vitamin E.

example 1

Amidated low-methoxylated pectin having a degree of methoxylation of 23 and a degree of amidation of 24 was dissolved in deionized water (4 g / 100 ml) to which various doses of human insulin (6, 15, 30 and 60 μ) were added and mixed using a blender ( Heidolph laboratory mixer, Germany) were stirred. Subsequently, dimethyl sulfoxide (3 ml) and vitamin E (3 ml) were added. This solution was mixed for a period of 6 (six) hours. Subsequently, an aliquot of the mixture (10 ml) was transferred to a petri dish (424.62 cm ² ) and frozen at -5 ° C. After freezing, a 2% CaCl ₂ solution was added to the frozen pectin and allowed to stand at room temperature for 10 minutes to allow cross-linking and formation of the matrix patch. Plasters with measured widths were cut out and applied to hydrofilm, which acted as a substrate. The patches were stored in a refrigerator at 2 ° C until use.

Example 2

The following variants of the method of Example 1 were carried out.

Example 2.1

• Drugs: NovoRapid insulin (NovoRapid FlexPen, Novo Nordisk, Canada)
• Matrix: Amidated, low-methoxylated pectin with a degree of methoxylation of 19 and a degree of amidation of 31.
• Cations for crosslinking: calcium chloride
• Penetration enhancer: dimethyl sulphoxide
• Adhesives: Self-adhesive bandages
• Antioxidants: vitamin E, eucalyptus oil
• Antibiotic: puromycin

Example 2.2

• Medicines: human insulin (NovoRapid FlexPen, Novo Nordisk, Canada)
• Matrix: Amidated, low-methoxylated pectin with a degree of methoxylation of 19 and a degree of amidation of 31.
• Cations for crosslinking: calcium chloride
• Penetration enhancer: sodium oleate
• Adhesives: Self-adhesive bandages
• Antioxidants: vitamin E, eucalyptus oil
• Antibiotic: puromycin

Example 2.3

• Medicines: human insulin (NovoRapid FlexPen, Novo Nordisk, Canada)
• Matrix: Amidated, low-methoxylated pectin with a degree of methoxylation of 19 and a degree of amidation of 31.
• Cations for crosslinking: calcium chloride
• Penetration enhancer: sodium dodecyl sulfate (SDS or NaDS)
• Adhesives: Self-adhesive bandages
• Antioxidants: vitamin E, eucalyptus oil
• Antibiotic: puromycin

Example 2.4

• Medicines: human insulin (NovoRapid FlexPen, Novo Nordisk, Canada)
• Matrix: Amidated, low-methoxylated pectin with a degree of methoxylation of 19 and a degree of amidation of 31.
• Cations for crosslinking: calcium chloride
• Penetration enhancer: dimethyl sulfoxide, sodium oleate
• Adhesives: Self-adhesive bandages
• Antioxidants: vitamin E, eucalyptus oil
• Antibiotic: puromycin

Example 2.5

• Medicines: human insulin (NovoRapid FlexPen, Novo Nordisk, Canada)
• Matrix: Amidated, low-methoxylated pectin with a degree of methoxylation of 19 and a degree of amidation of 31.
• Cations for crosslinking: calcium chloride
• Penetration enhancer: dimethyl sulfoxide, sodium dodecyl sulfate (SDS or NaDS)
• Adhesives: Self-adhesive bandages
• Antioxidants: vitamin E, eucalyptus oil
• Antibiotic: puromycin

Example 2.6

• Medicines: human insulin (NovoRapid FlexPen, Novo Nordisk, Canada)
• Matrix: Amidated, low-methoxylated pectin with a degree of methoxylation of 19 and a degree of amidation of 31.
• Cations for crosslinking: sodium dodecyl sulfate (SDS or NaDS), sodium oleate
• Adhesives: Self-adhesive bandages
• Antioxidants: vitamin E, eucalyptus oil
• Antibiotic: puromycin

Example 3

Amidated, low-methoxylated pectin having a degree of methoxylation of 19 and a degree of amidation of 31 was dissolved in deionized water (4 g / 100 ml) with stirring by use of a mixer (Heidolph Laboratory Mixer, Germany). Subsequently, either dimethyl sulfoxide, SDS or sodium oleate was added to the mixture. Vitamin E, eucalyptus oil and puromycin were added and mixed with stirring for 30 minutes. Various doses of NovoRapid insulin (4, 6, 8 and 10 units) were added to the mixture during the last 15 minutes of the preparation. Subsequently, an aliquot of the mixture (11 ml) was transferred to a petri dish (424.62 cm ² ) and frozen at -5 ° C. After freezing, the frozen pectin was allowed to stand at room temperature for 15 minutes and then a 2% CaCl ₂ solution was added to allow cross-linking and formation of the matrix patch. The patches were stored in a refrigerator at 2 ° C until use.

Transdermal delivery of insulin

animals

Male Sprague-Dawley rats (90-300 g body weight) were used, bred and maintained by the Institute of Biomedical Research of the University of KwaZulu-Natal. The animals had free access to standard rat chow (Meadows, Pietermaritzburg, South Africa) and water with a 12 hr light and 12 hr dark cycle. Procedures involving the animals and their care were carried out in accordance with the guidelines of the Kwa-Zulu Natal University (Ethical Clearance 007/10 / Animals).

In this study, male Sprague-Dawley rats (250-300 g), housed in the Institute for Biomedical Animal Resources at the university's Westville campus, were used by Kwa-Zulu Natal.

Determination of the amount of active ingredient in the patches

To determine the amount of drug incorporated in the patch, insulin content was determined in patches from known ranges. Plasters containing various doses of insulin were dissolved at pH 7.2 in Sorenson phosphate buffers. The amounts of insulin added to the Petri dishes for each group were 0.6, respectively; 1.5; 3.0 and 6.0. This corresponded to a theoretical amount of 0.027; 0.08; 0.135 and 0.27 of insulin added to each patch. Individual plasters were dissolved in the buffer and serial dilutions were made to measure the amount of insulin incorporated into each patch.

Application of hydrogel patch

The rats were shaved on the dorsal areas of the neck for 1-2 days prior to administration by the insulin patches. The hydrocapacity of the insulin hydrogel matrix patch was cut to the size of the patch and applied to an adhesive to allow easy transfer to the animal. The patches were held in place by adhesive hydrofilm (Hartman-Congo Inc, Rock Hill, South Carolina, USA) adapted to the sizes of the animals ( ).

Induction of experimental diabetes mellitus

Diabetes mellitus was induced in rats with a single intraperitoneal injection of streptozotocin (STZ, 60 mg / kg) dissolved in freshly prepared 0.1 M citrate buffer (pH 6.3). The control animals were injected with the vehicle. Control animals that showed glucosuria after 24 hours, tested by urine strips (Rapidmed Diagnostics, Sandton, South Africa), were considered diabetic. The blood glucose concentration of 20 mmol / L or over, measured after one week, was considered to be a stable diabetic condition before the experimental procedures were performed.

Experimental design

Non-diabetic and STZ-induced diabetic rats were divided into different groups for oral glucose tolerance (OGT) activity studies (n = 6 in each group).

Hydrogel insulin patch

OGT effects of amidated, hydrogel, pectin, insulin Matrix slides were studied in separate groups of non-diabetic and STZ-induced diabetic groups of rats in which the patch was applied to the shaved area on the back of the neck on the skin ( ). The control animals were apparently treated with drug-free pectin patches.

Oral glucose tolerance (OGT) reactions

OGT responses were evaluated in separate groups of non-diabetic and STZ-induced diabetic groups of rats in which the patch was applied to the shaved area on the back of the neck on the skin ( ). The rats were divided into the following groups: non-diabetic control animals, non-diabetic treated animals, STZ-induced diabetic control animals and STZ-induced diabetic treated rats (n = 6 in each group). Briefly, separate groups of non-diabetic and STZ-induced diabetic rats were eutrophicated overnight (18 hrs) followed by measurement of blood glucose (time 0). Subsequently, OGT responses to topically applied insulin, pectin, hydrogel patches were monitored at various insulin dosages (0.06, 0.21, 0.32 and 0.73 Lig · Kg ^-1 bwt). In the control group of animals, there was a sham application of drug-free pectin, hydrogel, matrix patches. Blood glucose was measured (with a blood glucose meter (Bayer's Glucometer Elite ^® (Elite Pty) Ltd, Healthcare Division, Isando, South Africa), before the glucose load and at 30, 60, 120 and 180 and 240 minutes after glucose load.

Determination of plasma insulin

The rats were sacrificed 4 hours after the start of the oral glucose tolerance test by an inhalation dose of halothane in an anesthesia chamber. Blood samples were then collected by cardiac puncture and transferred to heparinized tubes which were centrifuged immediately at 4 degrees Celsius and 3000 rpm for 15 minutes to pellet the blood cells. The supernatant (plasma) was aspirated with a Pasteur pipette. Plasma insulin concentrations were measured by ultra sensitive rat insulin ELISA tests evaluated (DRG Instruments GmBH, Marburg, Germany) with 100% cross-reactivity with insulin lispro (Humalog ^® Eli Lilly). The immunoassay is a quantitative method for the determination of plasma insulin using two monoclonal antibodies which together are specific for insulin. The lower detection limit was 1.74 pmol ^-1 . The intra- and inter-assay analytical coefficients of variation ranged from 4.4% to 5.5% and 4.7% to 8.9%, respectively.

Statistical analysis

All data were expressed as mean ± standard error of the device (S.E.M). Statistical comparison of differences between control means and experimental group was made by GraphPad InStat Software (Version 4.00, GraphPad Software, San Diego, Calif.) Using a one-way variance analysis (ANOVA), after Tukey-Kramer multiple comparison, followed by Tukey -Kramer multiple comparison test. A value of p <0.05 was considered significant.

Results

dissolution studies

Table 2 shows the amount of insulin in insulin pectin hydrogel patches. The theoretical amount of insulin in each patch was determined by the known amount of insulin added to the Petri dishes during the patch preparation and the area of the plasters cut from the Petri dishes. The insulin incorporation in each patch ranged from 70% to 81%. Table 2 Results of insulin patch hydrogel pectin dissolution studies Insulin in Petri dish (μg) Theoretical insulin in plaster (μg) Actual insulin in patch (μg) Dosage (μg) · kg- ¹ b · wt % Insulin incorporation Low 0.6 0.027 0.019 0.06 70 Int. low 1.5 0.08 0.062 0.21 75 Int. high 3.0 0.135 0,095 0.32 70 High 6.0 0.27 0.22 0.73 81

Glucose tolerance reactions

shows the blood glucose responses of 5 groups of diabetic rats (n = 10) to an oral glucose load. The 5 different groups were untreated control animals, as well as rats with 0.06; 0.21; 0.32 and 0.73 | (μg) · kg ^-1 insulin were treated by a pectin hydrogel patch. To simplify the test results, the four treatment groups were each referred to as low dosage, medium dosage, higher dosage and high dosage.

Treatment with the high dose of insulin resulted in a significantly (p <0.05) lower blood glucose concentration at all times throughout the glucose tolerance test compared to all other insulin doses. For the 2 lowest doses, no significant difference (p> 0.05) was seen in the blood glucose reactions at all time points compared to the control group. In the rats treated with the medium dose, a significant reduction (p <0.05) of the blood sugar was perceived, in comparison with the control group and 2 lower dose groups.

Plasma insulin concentrations

The plasma insulin concentrations of streptozotocin (STZ) -induced diabetic rats treated with various doses of insulin in a pectin hydrogel patch measured 4 hours after the onset of the glucose effect test are shown in , The values are represented as mean values and vertical bars show the SEM of the mean values (n = 6 in each group). * p <0.05 when compared with control animals.

No statistically significant difference (p> 0.05) in plasma insulin concentrations between the low dose and the control dose. The plasma insulin concentrations were significantly (p <0.05) higher in all animals compared to the control animals. The plasma insulin concentrations in the high-dose insulin-treated animals were significantly higher (p <0.05) than in any other group.

discussion

The invention relates to an adherent pectin hydrogel skin patch, which can deliver insulin into the bloodstream, while reducing the plasma glucose concentration in STZ-induced diabetic rats. Transdermal drug delivery is non-invasive, allowing slow controlled release of drugs and reducing gastric and hepatic metabolism. Drug formulations from the pharmaceutical industry have previously consisted of simple, fast-acting chemical compounds delivered orally or by injection. The use of transdermal drug delivery is usually limited by a low permeability of the skin, and the present invention demonstrates the improved permeability of drugs through the skin. Insulin is used extensively and the worldwide spread of diabetes mellitus represents a huge market potential. About 215 million people currently suffer from diabetes mellitus. The treatment of diabetes usually requires daily subcutaneous (sc) injections, and therefore, transdermal insulin delivery frees patients from daily injections, thereby improving patient adherence. The major difference between the pectin pilaster of the invention and prior transdermal delivery systems is that the pilaster of the invention has the ability to transport insulin through the skin without the use of additional mechanisms.

literature

• 1. Stevring H, Andersen M, Beck-Nielsen H, Green A and Vach W. (2007) Counting drugs to understand the disease: The case of measuring the diabetes epidemic. Population Health Metrics, 5: 2.
• 2. Musabayane CT, Munjeri O, Bwititi P and Osim EE. (2000). Orally insulin-administered, insulin-loaded amidated pectin hydrogel beads sustain plasma concentrations of insulin in streptozotocin-diabetic rats. Journal of Endocrinology: 164: 1-6.
• 3. Transdermal insulin delivery using lipid enhanced electroporation. Sen, Daly and Hui. Biochimica et Biophysica Acta (BBA) biomemebranes. 1564 (1), August 2002, 5-8.
• 4. Transdermal delivery of insulin from a novel biphasic lipid system in diabetic rats. King, MJ; Badea, I; Solomon J; Kumar, P .; Gaspar, KJ and Foldvari, M. Diabetes Techol Ther. 4 (4), 2002: 479-88.
• 5. Transdermal drug delivery of insulin with ultra-deformable carriers. Cevc, G. Clinical Pharmacokinetics. 2003; 42 (5): 461-74.
• 6. Noninvasive ultrasonic transdermal insulin delivery in rabbits using the lightweight cymbral array. Lee, S; Snyder, B; Newnham, RE and Smith NB. Diabetes Technology Therapy. 2004, 6 (6): 808-15.
• 7. Transdermal delivery of insulin using microneedles in vivo. Martano, W; Davis, SP; Holiday, NR; Wang, J; Gill, HS and Prausnitz, MR. Pharmacology Research 2004 21 (6), 947-952
• 8. Transdermal delivery of regular insulin to chronic diabetic rats: effect of skin preparation and electrical enhancement. Zakzewski, CA; Wasilveski, J; Cawley, P and Ford, W. Journal of Control Release. 1998, 50 (1-3): 267-272.

Claims (8)

A transdermal delivery system for the transdermal delivery of insulin, in the form of a transdermal patch comprising: cross-linked, amidated, low-methoxylated pectin; Insulin; and a transdermal transfer accelerator. The transdermal delivery system of claim 1 wherein the amidated low methylated pectin has a degree of methoxylation of about 19 to 23 and a degree of amidation of about 24 to about 31. The transdermal delivery system of claim 1 or claim 2, wherein the transdermal delivery enhancer is selected from dimethylsulfoxide, sodium oleate, sodium dodecylsulfate, or a combination of two or more thereof. Transdermal delivery system according to any one of the preceding claims, wherein the patch contains an antioxidant. Transdermal administration system for transdermal administration according to claim 4, wherein the antioxidant is vitamin E. Transdermal delivery system according to claim 4 or claim 5, wherein the antioxidant is vitamin E in combination with eucalyptus oil. A transdermal delivery system according to any one of the preceding claims, wherein the patch further contains an antibiotic. A transdermal delivery system for administration according to claim 7, wherein the antibiotic is puromycin (purmycin).

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